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Applied Biosystems™ TaqMan™ hPSC Scorecard™ Panel, Fast 96-well

Catalog No. A15876
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Description

TaqMan™ hPSC Scorecard™ Panel, Fast 96-well

The Applied Biosystems™ TaqMan™ hPSC Scorecard™ Panel 2 x 96w FAST enables verification of pluripotency and determination of lineage bias for both ES and iPS cell lines. Each of the two 96-well plates contains 94 predefined TaqMan™ Gene Expression assays (including endogenous controls) dried-down in the wells. Master mix is not included with this panel. For a kit that combines the panel with master mix, see TaqMan™ hPSC Scorecard™ Kit 2 x 96w FAST. This product is also available in a 384-well format.

  • Assess an entire gene signature in one simple experiment
  • Compare results to a reference standard with the accompanying hPSC Scorecard™ Analysis Software
  • Identify lines with trilineage differentiation potential
  • Evaluate inductive differentiation specified to one germ layer
  • Access validated content for increased confidence in your results

    Assess an Entire Gene Signature in One Experiment
    The TaqMan™ hPSC Scorecard™ Panel helps save time by offering pre-plated assays in a stable and convenient dried-down format. Simply add TaqMan™ Master Mix to your cDNA of interest and transfer to the 96-well plate with a multichannel pipette.

    Compare Results to a Reference Standard
    The TaqMan™ hPSC Scorecard™ Panel includes access to proprietary cloud-based analysis software that allows you to view and export graphs and tables of results, as well as generate reports. The software is compatible with seven Applied Biosystems™ qRT-PCR systems and is available at no additional charge.

    Identify Lines with Trilineage Differentiation Potential
    Include randomly differentiated embryoid body (EB) samples (differentiated for at least 7 days using standard EB methods) in your analysis and determine whether your lines are biased toward one or more of the three germ layers (endoderm, mesoderm, ectoderm).

    Evaluate Inductive Differentiation Specified to One Germ Layer
    The panel includes over twenty markers for each of the three germ layers that have been shown to respond as expected to directed differentiation in monolayer culture. Quickly evaluate time course data, culture conditions and media formulations for your germ layer of interest with this one panel.

    Access Validated Content
    The panel contents are based on published work (Bock et al., Cell 144, 439–452, 2011) and were validated against multiple human ES and iPS lines.

    What You Get
    The TaqMan™ hPSC Scorecard™ Panel 2 x 96w FAST includes:

  • Two 96-well plates of 94 TaqMan™ Gene Expression assays
  • Two optical plate covers

    • Specifications

    Type Panel
    Format 96-array Plate
    Species Human
    Content And Storage Contents: Two 96-well plates each containing the 96-assay panel and two optical covers

    Store at room temperature.
    Detection Method Primer-Probe Detection
    Form Dried Down
    For Use With (Equipment) QuantStudio™ 12k Flex, StepOne™, Fast Mode, ViiA™ 7 System
    Includes 2 x 96-well plate
    Product Line TaqMan
    Quantity 2 x 96-well plates
    Reaction Speed Fast
    Research Category Stem Cell Research
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    Frequently Asked Questions (FAQs)

    I'm using the TaqMan hPSC Scorecard Panel. How do I load the samples onto a 384-well plate if I don't have a multichannel pipette?

    You can load samples using a single channel pipette, but this method is time-consuming and may increase pipetting error. We strongly recommend that you use an 8- or 16-channel multichannel pipette. You also can dispense samples using automated systems, with the understanding that additional sample will be required to compensate for the dead volume.
    Why should I set up cDNA synthesis in 8 wells of a 96-well plate or in 8-well PCR strips when using the TaqMan hPSC Scorecard Panel?

    Setting up your cDNA synthesis in 8 wells of a 96-well plate or in 8-well PCR strips facilitates sample loading of the 96-well and 384-well hPSC Scorecard Panel plates.
    Can I analyze somatic non-pluripotent primary cells with the TaqMan hPSC Scorecard Panel?

    Somatic non-pluripotent primary cells, such as the parental lines used for iPSC generation, are not pluripotent in nature and their scores will be low. However, the expression of lineage markers will largely rely on homogeneity of the cells. Note that the markers in the panel are designed to evaluate early germ layer specification and not any particular terminally differentiated state.

    Does the reprogramming method affect the TaqMan hPSC Scorecard analysis results?

    Minor differences in gene expression profiles are sometimes observed based on the reprogramming method, but in general the hPSC Scorecard analysis results do not change significantly for lines derived using various reprogramming methods. This was tested with ESC and iPS derived using episomal or Sendai-based reprogramming systems before and after seven days of spontaneous differentiation. Cells were grown in KSR-based media on irradiated MEFs prior to removal of FGF for EB formation.

    Will the presence of Sendai virus affect my TaqMan hPSC Scorecard analysis results?

    The presence or absence of Sendai virus in established iPSC clones does not have an impact on pluripotency and hence on TaqMan hPSC Scorecard analysis results.

    When I use Sendai virus (SEV) to generate iPSCs for TaqMan hPSC Scorecard analysis, can I check for the presence of residual virus?

    Sendai virus (SEV) is included in the TaqMan hPSC Scorecard kit and can detect the presence of the Sendai virus backbone. Note that this method will not distinguish between the different reprogramming factors but just the presence or absence of the residual virus in the cells. If an unexpected signal is detected, check the amplification curve to determine if it is a false positive. If it's a false positive, you may ignore the flag.

    For TaqMan hPSC Scorecard analysis, what is the earliest passage that I can use when I generate iPSCs?

    We recommend that you culture iPSC clones to at least passage 8-10 until they are stable and homogeneous, prior to hPSC Scorecard analysis. Early passage iPSC clones may give low self-renewal ("pluri" in v1.1 of the analysis software) scores or show higher expression of lineage genes.

    Have you tested cells grown in Gibco PSC Neural Induction Medium (NIM) for Scorecard analysis?

    H9 cells were differentiated into NSCs using Gibco PSC Neural Induction Medium (NIM) and hPSC Scorecard analysis was performed at various time points. The control sample was undifferentiated H9 ESC. Cells were seen to become positive for ectoderm by day 5.

    I am planning to use the TaqMan hPSC Scorecard kit/panel. I am only interested in directed differentiated of cells. Do I need to use a particular method for directed differentiation?

    You can perform directed differentiation according to your own methods. However, the time point when expression is noticeable will largely depend on the robustness of the methods. We recommend testing a few time points to monitor differentiation with time.

    I am planning to use the TaqMan hPSC Scorecard kit/panel. What is the minimum time required to form embryoid bodies (EBs) and is there a specific protocol?

    You can differentiate cells using any of the established methods. When using suspension embryoid bodies, we recommend that you allow at least 7 days for differentiation prior to analysis.
    For the TaqMan hPSC Scorecard kit/panel, can I use cells on a feeder-free system cultured in novel media?

    The TaqMan hPSC Scorecard kit/panel measures self-renewal and trilineage differentiation potential of PSCs and is not restricted by culture conditions. You should be aware that using novel media systems may have particular effects on the cells in terms of pluripotency and/or their differentiation. We recommend designing experiments that use novel media by including a control condition that utilizes traditional media or media in which expression patterns have been tested and confirmed. The most recent version of the hPSC Scorecard Analysis application offers a differentiation index plot for viewing gene expression in embryoid bodies (EBs) relative to the undifferentiated state.

    With the TaqMan hPSC Scorecard kit/panel, how do I harvest human PSCs cultured using feeder-dependent culture systems?

    We recommend removing feeders from your culture before proceeding with Scorecard analysis. To remove feeders, harvest your PSCs with collagenase, allow the colonies to settle by gravity sedimentation to reduce feeder-carryover, and re-seed cells in feeder-free conditions on Geltrex matrix-coated dishes and MEF conditioned medium for 1 to 2 passages. The presence of feeders can contribute to gene expression measured in the assay, thus altering the gene-signature pattern.

    For the TaqMan hPSC Scorecard kit/panel, can I use cells that are on feeders or in feeder-free conditions?

    Yes. The TaqMan hPSC Scorecard kit/panel measures the potential for self-renewal and trilineage differentiation of PSCs grown on feeders or in feeder-free conditions.
    How many cells are required to run one experiment with each TaqMan hPSC Scorecard kit/panel?

    While we recommend that you use about 0.5 million cells per experiment, or the number of cells equivalent to 1 well of a 6-well dish, you can use as few as 100,000 cells when performing RNA purification. If you perform a lysis protocol using Cells-to-CT or CellsDirect One-Step qPCR kits, as few as 15,000 cells is sufficient. Please note, however, that reducing cell number can compromise the quality of the results.
    What is included with each TaqMan hPSC Scorecard kit or panel?

    Panel configurations:
    - Two 96-well (2 x 96w FAST) plates with optical plate covers
    - One 384-well (384w) plate with optical plate covers

    Kit configurations:
    - Two 96-well (2 x 96w FAST) plates with optical plate covers & TaqMan Gene Expression Master Mix
    - One 384-well (384w) plate with optical plate covers & TaqMan Gene Expression Master Mix

    For Research Use Only. Not for use in diagnostic procedures.